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Waters 1500-Series, Operator'S Manual

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Wavelength selection

C-15

Ethanol

1.361

Cyclohexane

1.427

Methyl acetate

1.362

Ethylene glycol

1.427

Isopropyl ether

1.368

N,N-

Dimethyl formamide 

(DMF)

1.428

Ethyl acetate

1.370

N,N-

Dimethyl acetamide 

(DMAC)

1.438

1-Pentene

1.371

Ethyl sulfide

1.442

Acetic acid

1.372

Chloroform

1.443

Isopropyl chloride

1.378

Ethylene dichloride

1.445

Isopropanol

1.38

Carbon tetrachloride

1.466

n

-Propanol

1.38

Dimethyl sulfoxide 

(DMSO)

1.477

Methylethylketone

1.381

Toluene

1.496

Diethyl amine

1.387

Xylene

~1.50

n

-Propyl chloride

1.389

Benzene

1.501

Methylisobutylketone

1.394

Pyridine

1.510

Nitromethane

1.394

Chlorobenzene

1.525

1-Nitropropane

1.400

o

-Chlorophenol

1.547

Isooctane

1.404

Aniline

1.586

Cyclopentane

1.406

Carbon disulfide

1.626

Refractive indices for common chromatographic solvents (Continued)

Solvent

RI

Solvent

RI

Summary of Contents for 1500-Series

Page 1: ...Waters 1500 Series HPLC Pump and Options Operator s Guide 715002013IVD Revision C Copyright Waters Corporation 2014 All rights reserved ...

Page 2: ...d accurate at the time of publication In no event shall Waters Corporation be liable for incidental or consequential damages in connection with or arising from its use Trademarks Waters is a registered trademark of Waters Corporation and Breeze Empower LAC E SAT IN and THE SCIENCE OF WHAT S POSSIBLE are trademarks of Waters Corporation Luer is a registered trademark of Becton Dickinson and Company...

Page 3: ...r disposal of any Waters product You can reach us via the Internet telephone or conventional mail Safety considerations Some reagents and samples used with Waters instruments and devices can pose chemical biological and radiological hazards You must know the potentially hazardous effects of all substances you work with Always follow Waters contact information Contacting medium Information Internet...

Page 4: ...00 V Back siphoning and draining Check valves in the fluid pump installed ahead of the column heater prevent the back siphoning of fluids Drainage systems are installed in this equipment Drip trays inside the pump and column heater units catch any fluid from leaks or spills These trays are connected to external drains on the bottom of the units Tubing connected to this drain routes the fluid into ...

Page 5: ...Europe Safety advisories Consult Appendix A for a comprehensive list of warning and caution advisories Operating the Waters 1500 Series HPLC pump and options When operating these pumps and options follow standard quality control QC procedures and the guidelines presented in this section ...

Page 6: ...he optional 1500 Series Column Heater and its options to deliver a precisely controlled amount of solvent to a column maintaining a consistent and reproducible mobile phase composition The Waters 1500 Series HPLC pump and options are for research use only Symbol Definition Manufacturer Authorized representative of the European Community Confirms that a manufactured product complies with all applic...

Page 7: ...l within an acceptable range and evaluate precision from day to day and run to run Data collected when QC samples are out of range might not be valid Do not report these data until you are certain that the instrument performs satisfactorily When analyzing samples from a complex matrix such as soil tissue serum plasma whole blood and other sources note that the matrix components can adversely affec...

Page 8: ...or used conductively coupled radio frequency energy that is necessary for the internal functioning of the equipment Class B products are suitable for use in both commercial and residential locations and can be directly connected to a low voltage power supply network EC authorized representative Waters Corporation Stamford Avenue Altrincham Road Wilmslow SK9 4AX United Kingdom Telephone 44 161 946 ...

Page 9: ...rs 1500 Series HPLC pump and options vi Calibrating vi Quality control vi ISM classification vii ISM Classification ISM Group 1 Class B vii EC authorized representative viii 1 Introduction 1 1 HPLC pump operating principles 1 2 Isocratic and gradient LC system operation 1 2 Effects of dissolved oxygen on the mobile phase 1 3 Using in line degassing to remove gases from eluents 1 4 Overview of the ...

Page 10: ...EE 488 connections 2 6 Setting the IEEE 488 address 2 9 Connecting pump inlet and outlet lines 2 11 Connecting the eluent supply 2 11 Connecting the pump outlet 2 14 Connecting fluid waste lines 2 16 3 Installing Options and Accessories 3 1 Installing the 1500 series column heater 3 2 Installing a manual injector 3 4 Connecting to the column or column heater 3 5 Connecting the inject start signal ...

Page 11: ... integral vacuum degasser 4 5 Operating with the plunger seal wash system 4 8 Maximum flow rates for 1500 series pumps 4 10 Preparing for Breeze 2 operation 4 10 Priming and purging the pump via Breeze 2 control 4 11 Purging the flow path via Breeze 2 control 4 12 Equilibrating the system via Breeze 2 control 4 13 Preparing for Empower 2 operation 4 15 Priming and purging the pump via Empower 2 co...

Page 12: ... replacing the plungers 5 9 Replacing check valves 5 14 Replacing 1515 1525 check valves 5 15 Replacing 1525EF check valves 5 17 Replacing 1525µ check valves 5 19 Replacing a draw off valve 5 22 Removing the draw off valve 5 23 Installing a draw off valve 5 24 Replacing fuses 5 25 Replacing the rear panel fuses 5 26 6 Troubleshooting 6 1 Troubleshooting pump problems 6 2 Identifying and correcting...

Page 13: ...klist C 2 Water C 2 Buffers C 3 Tetrahydrofuran THF C 3 Solvent compatibility C 3 Solvents to avoid C 3 Solvents to use C 3 Solvent miscibility C 6 How to use miscibility numbers M numbers C 8 Buffered solvents C 8 Head height C 9 Solvent viscosity C 9 Mobile phase solvent degassing C 9 Gas solubility C 9 Eluent degassing methods C 10 Wavelength selection C 12 UV cutoffs for common solvents C 12 M...

Page 14: ...xiv Table of Contents ...

Page 15: ... data control configurations and available options for the 1500 series HPLC pumps Contents Topic Page HPLC pump operating principles 1 2 Overview of the 1500 series HPLC pumps 1 5 Fluid handling components 1 7 Electronic components 1 12 Pump control 1 13 Options and accessories 1 15 ...

Page 16: ... proceeds the elution strength of the mobile phase is increased to elute the more strongly retained sample components In the simplest configuration there are two bottles of solvent and two pumps the case when using a 1525 binary pump The speed of each pump is managed by the gradient controller to deliver more or less of each solvent over the course of the separation The two streams are combined us...

Page 17: ...nd irregularity Removing dissolved oxygen to a reproducible level greatly enhances the performance of UV Vis detectors especially below 254 nm and in gradient systems It also improves sensitivity in certain fluorescence detection applications Effects on fluorescence detectors For certain analytes at certain wavelengths oxygen under certain mobile phase conditions can quench fluorescence response A...

Page 18: ...ws the relationship between the flow rate of an eluent water and the concentration of a gas oxygen dissolved in the eluent Degasser operating principles The degasser operates according to Henry s Law removing dissolved gases from the eluent Henry s Law states that the mole fraction of a gas dissolved in a liquid is proportional to the partial pressure of that gas in the vapor phase above the liqui...

Page 19: ...g gas Surface area of degassing membrane The length of the degassing membrane is fixed in each vacuum chamber Overview of the 1500 series HPLC pumps The 1500 series HPLC pumps combine the most important aspects of eluent delivery for HPLC high precision reliability and smooth eluent flow All pumps perform their intended functions equally well The 1515 isocratic HPLC pump is designed for precise is...

Page 20: ...ing or eluent compressibility These optional components are available for the 1500 series pump to suit your HPLC applications and site requirements 1500 series column heater Enables preheating of fluids passing through chromatographic columns Integral vacuum degasser Provides HPLC pumps with an automatic continuous method of removing dissolved gasses from mobile phases The degasser is standard on ...

Page 21: ...elf with its fluid handling components The following figures identify the fluid handling components of the 1525 1525EF and 1525µ HPLC pumps Tip The 1515 pump is an isocratic version of the 1525 binary pump It shares key components with the 1525 binary pump except for the second pump assembly Pump B tee and gradient mixer including associated subcomponents ...

Page 22: ...head valve assembly assembly manifold Draw off Gradient mixer Pulse dampeners covered by shroud Pressure Tee transducer Pump outlet Inlet check valve assembly Pump B Pump A Indicator rod Reference valve waste line Manual injector waste lines Drip tray Drip tray waste exit valve valve Manual injector optional ...

Page 23: ...mixer Inlet manifold Restrictor tee Pressure transducer Pump head assembly Draw off valve Drip tray waste exit Drip tray Integral vacuum Solvent inlets Vent valve Inlet check valve assembly Outlet check valve assembly Indicator rod Pulse dampeners behind shroud Pump outlet Vent valve waste line degasser ...

Page 24: ...ening in one direction only Inlet manifold Provides the connection for eluent inlet tubing and routes eluent to the inlet check valve on each pump head Integral vacuum degasser optional for 1515 and 1525 pumps Provides HPLC pumps with an automatic continuous method of removing dissolved gasses from mobile phases Manual injector optional Enables a manual sample injection Generates an inject start s...

Page 25: ... injector the column and the rest of the system Reference or vent valve waste line Routes flow from the reference or vent valve to the waste container Restrictor tee 1525µ pump only Blends two solvents with backflow prevention Seal wash holes not visible Allow manual flushing of plunger seals Tee 1525 1525EF pump only Blends two solvents Fluid handling components Continued Component Description ...

Page 26: ...of the 1500 series pump s electronic components Electronic components in a 1500 series pump Component Description Cooling fan vent Exhausts air for cooling internal electronics I O terminal block Provides input and output contact closures for connecting to external devices Ethernet port Power cord Power switch Fuse holder side of unit I O terminal block IEEE 488 port IEEE 488 address switches Cool...

Page 27: ...nd optional column heater communicate with the data system via an Ethernet communications interface Where all system components including a 1500 series pump and optional column heater communicate with the data system via an IEEE 488 bus interface You cannot use the pump or column heater s Ethernet port at the same time you are using its IEEE 488 bus interface for communications Power entry module ...

Page 28: ... more information see page 2 5 HPLC system configuration using Ethernet communications IEEE 488 configuration To communicate with the Waters data control system via IEEE 488 an IEEE 488 cable connects the 1500 series pump to an IEEE 488 controller a busLAC E card in the data control system for Breeze or Empower control or to a NI GPIB card for MassLynx control For more information see page 2 6 Eth...

Page 29: ... the pump for easy access see the figure 1525 binary pump with optional column heater on page 1 7 For installation instructions see page 3 2 Integral vacuum degasser The integral vacuum degasser is a standard feature of the 1525µ binary pump and an optional feature of the 1515 1525 1525EF HPLC pumps It provides HPLC systems with an automatic continuous method of removing dissolved gases from mobil...

Page 30: ...s except the 1525µ Ιt consists of a single injector valve that you can install and use to perform precise manual sample injections The FlexInject manual dual injector operates with any fluid mixing system including 1500 series pumps It allows you to inject small or large sample volumes by simply selecting a valve The FlexInject injector can operate with flow rates as high as 150 mL min and mounts ...

Page 31: ...thernet Instrument Getting Started Guide Gradient mixers The Waters 1500 series pumps support various configurations of gradient mixers that vary in eluent volume capacity You can order and install one or more of these mixers in a 1500 series pump For installation instructions see page 3 8 ...

Page 32: ...1 18 Introduction ...

Page 33: ...guration includes options such as a column heater or accessories such as a different eluent mixer refer to the appropriate sections in Chapter 3 to complete your 1500 series pump installation Contents Topic Page Site requirements 2 2 Unpacking 2 3 Connecting power and signal cables 2 4 Connecting pump inlet and outlet lines 2 11 ...

Page 34: ...ter Depth 61 cm Height 43 cm Vibration Negligible Clearance At least 15 cm at rear for ventilation and cable connections Static electricity Negligible Input voltage and frequency Grounded AC 120 240 VAC 50 60 Hz single phase Electromagnetic fields No nearby source of electromagnetic noise such as arcing relays or electric motors Power requirement 200 VA for pump 150 VA for column heater Caution To...

Page 35: ...ters 1500 series HPLC pump is shipped in a single carton that contains the following items Waters 1515 Isocratic 1525 Binary or 1525µ HPLC Pump Startup kit TP01723 61 cm 30 5 cm 43 cm 15 cm 12 7 cm Clearance for optional column heater Clearance for bottle holder ...

Page 36: ...ation with this guide for future reference 6 Inspect all items for damage and immediately report any shipping damage to both the shipping company and your Waters representative If shipping damage occurred contact Waters Customer Service Refer to Waters Licenses Warranties and Support for complete information on shipping damages and claims Connecting power and signal cables For proper operation the...

Page 37: ...ounded power outlet Making ethernet connections The 1500 series pump other chromatography equipment in the HPLC system such as the column heater autosampler detector and the data control workstation can interconnect using Ethernet cables and a networking switch see page 1 16 The pump is equipped with a RJ 45 connector for Ethernet port communications The Ethernet port a 10 100 Base T networking in...

Page 38: ...tions If you are not using Ethernet communications for your HPLC configuration the 1500 series pump interconnect other chromatography equipment in the HPLC system such as the column heater autosampler detector and the data control workstation using IEEE 488 cables Most chromatography equipment is shipped with a 1 meter cable with dual receptacle connectors at each end The Waters data control works...

Page 39: ...low these guidelines when you install and use your system Always keep all devices powered on while your system is in use While a system is active on the IEEE 488 bus do not power on or off any device on the bus The maximum number of devices that can be connected together to form one interface system is 15 14 instruments plus the busLAC E or NI GPIB card The maximum total cable length to connect th...

Page 40: ... receptacle end of the 2 meter IEEE 488 cable to the busLAC E card in the Empower or Breeze 2 workstation or to the NI GPIB card in the MassLynx workstation IHPLC system using IEEE 488 connections 2 Connect the dual receptacle end of the 2 meter IEEE 488 cable to the IEEE 488 port on one of the IEEE 488 devices pump column heater autosampler or detector 3 Connect one end of a 1 meter IEEE 488 cabl...

Page 41: ...ear panel of the pump from 0 to 1 see the table below For example to obtain an address of 7 add the numbers of the switches in the 1 position Setting the address switches To set the IEEE 488 address 1 Ensure the unit is powered off 2 Set the DIP switches on the rear panel of the pump to a unique IEEE 488 address referring to the following table for DIP switch settings for valid IEEE 488 addresses ...

Page 42: ...1 0 13 1 0 1 1 0 14 0 1 1 1 0 15 1 1 1 1 0 16 0 0 0 0 1 17 1 0 0 0 1 18 0 1 0 0 1 19 1 1 0 0 1 20 0 0 1 0 1 21 1 0 1 0 1 22 0 1 1 0 1 23 1 1 1 0 1 24 0 0 0 1 1 25 1 0 0 1 1 26 0 1 0 1 1 27 1 1 0 1 1 28 0 0 1 1 1 29 1 0 1 1 1 IEEE 488 switch settings Continued IEEE 488 address Switch settings 1 2 4 8 16 ...

Page 43: ...ing the degasser to the eluent reservoir and pump inlet Required materials Tefzel ferrule and compression screw startup kit one set per pump 1 8 inch OD ETFE tubing part number WAT270714 in startup kit ETFE tubing startup kit part number WAT024036 in startup kit Reservoir containing filtered degassed eluent one per pump Bottle holder Bottle caps 1 L part number WAT062479 in startup kit Stainless s...

Page 44: ...en using high viscosity eluents you can possibly need to increase reservoir height 46 to 61 cm above the pump s inlet manifold 2 Insert the ETFE tubing into the 1 8 inch diameter hole of the tubing cutter making sure that the tubing that extends from the metal side of the cutter is the correct length as determined in step 1 3 Insert the razor blade into the cutter and press down to cut the tubing ...

Page 45: ...lene containers as reservoirs When using the instrument in combination with a mass spectrometer refer to recommendations published in the most recent version of Controlling Contamination in UPLC MS and HPLC MS Systems on the Waters Web site Choose 1 L solvent reservoirs that provide a snug fit for the reservoir caps supplied in the startup kit The solvent reservoir caps help control solvent evapor...

Page 46: ... step 5 for the second pump assembly Tip If you are connecting the pump to a Waters 717plus or 2707 autosampler you can use one position of the bottle holder to hold a bottle that contains the needle wash solvent Connecting the pump outlet Follow the instructions in this section to connect the pump outlet to the injector or next component in the flow path Making pump outlet connections involves th...

Page 47: ...r instead of the knife edge file to obtain a clean square cut 3 Grasp the tubing on both sides of the scribe mark with cloth covered pliers to prevent marring of the tube surface and gently work the tube back and forth until it snaps Requirement Ensure that the break is square and free of burrs Attaching the compression fittings to the tubing To attach the fittings 1 Slide a compression screw onto...

Page 48: ... end of the outlet tubing disconnected until you have primed the pump as described in Chapter 4 2 After you primed the pump press the free end of the tubing assembly into the injector or next device in your HPLC system and finger tighten the compression screw 3 Use the 5 16 inch wrench to tighten the screw another 1 8 turn Connecting fluid waste lines Follow the instructions in this section to con...

Page 49: ...the 1525 1525EF pumps on page 1 8 or the figure Fluid handling components in the 1525µ pump on page 1 9 and secure it in the provided spring clips 4 Insert the reference valve or vent valve waste line and the optional manual injector waste lines together in the top end of the dual tubing 5 Insert the other end of the dual tubing in a properly situated waste container To connect the drip tray to th...

Page 50: ...2 18 Installing the HPLC Pump ...

Page 51: ...of 1500 series pump options and accessories as well as ordering information and prerequisites visit the Waters Web site or contact Waters Contents Topic Page Installing the 1500 series column heater 3 2 Installing a manual injector 3 4 Installing different eluent mixers 3 8 Installing the integral vacuum degasser 3 14 Installing the plunger seal wash system 3 17 ...

Page 52: ... column heater unit Ensure that users have unobstructed access to the column heater compartment The column heater does not contain any user serviceable parts Do not use tools to access the components inside the device To install the column heater 1 Remove the three screws from the pump s right hand side panel 2 Insert the screws through the standoffs included with the Column Heater Startup Kit so ...

Page 53: ...elocate the column heater after initial installation 1 Note the current location of all electrical and tubing connections 2 Power off the system and disconnect all electrical cables 3 Disconnect any tubing that directs flow through the column heater 4 Verify that the drip tray is empty and then disconnect the drip tray hose from the underside of the device 5 Carefully move the device to the new lo...

Page 54: ...u are installing a 1500 series manual injector or FlexInject dual manual injector refer to the installation instructions and startup list included with the injector Requirements If you are installing a manual injector you must connect the pump outlet and injector waste lines according to the instructions on page 2 14 and page 2 16 When using a manual injector you must configure an inject start sig...

Page 55: ...n the detector This section describes the connections for Waters 2489 and 2414 Detectors For a description of signal connections to other model detectors refer to the detector s operator s guide Required materials Two wire cable from the manual injector Screwdriver small flat blade To make the inject start signal connection from the pump to the detector 1 Remove the terminal block from the rear of...

Page 56: ...then tighten the screw to secure the wire Tip To connect the inject start pump signal to other detector inputs for example to the Autozero inputs refer to the appropriate detector operator s guide for details 8 Firmly push the terminal block into the back of the detector I O terminal block connections Function Pin number Direction Inject Start 1 Output or Input Inject Start 2 Stop Flow 3 Input Sto...

Page 57: ...n When you use the manual injector in conjunction with compositional gradients turn the injector back to the LOAD position before the end of the run Leaving the injector in the INJECT position automatically triggers the gradient table for subsequent injections but data acquisition does not start For instructions on operating the FlexInject Dual Manual Injector with a detector refer to the Waters F...

Page 58: ...position for a minimum of two loop volumes before turning back to the LOAD position to start acquiring data Tip For instructions on operating the FlexInject Dual Manual Injector with a detector refer to the Waters FlexInject Dual Manual Injector Module Installation Guide Installing different eluent mixers You can use various eluent mixer configurations in the 1500 series pumps This section describ...

Page 59: ... three configurations shown in the table above 1525 binary HPLC pump The 1525 pump is shipped with one standard GM150 gradient mixer part number WAT055847 mounted on the upper left hand side of the chassis face shown as configuration 1 in the figure below If you require a second GM150 mixer see configuration 2 If you want to replace the standard GM150 mixer with a large volume gradient mixer see c...

Page 60: ...led 1 2 and 3 Installing a single GM150 gradient mixer Tip This mixer is standard on the 1525 pump 1 2 3 Outlets Inlets Reference valve Pressure transducer Configuration 1 Single GM150 mixer Configuration 2 Dual GM150 mixers Configuration 3 Large volume mixer Oriented horizontally top to bottom ...

Page 61: ...xer outlet and the other end to the reference valve inlet using a compression screw and ferrule on each end To install the single GM150 gradient mixer on the 1525µ pump 1 Acquire a mixer part number WAT055847 and the mixer mounting bracket and 4 mounting screws startup kit 2 Loosen the compression screw and disconnect the tubing that connects the pressure transducer outlet to the 50 µl mixer inlet...

Page 62: ...kit and attach one end to the pressure transducer outlet and the other end to the bottom mixer inlet using a compression screw and ferrule on each end 6 Cut one 9 25 cm minimum length of 0 009 inch diameter tubing part number WAT026973 in the startup kit bend it to form a U tube and place a compression screw and ferrule on each end 7 Connect the outlet of the bottom mixer to the inlet of the top m...

Page 63: ... configured for a GM150 mixer perform these actions a Remove the tubing that connects the pressure transducer outlet to the GM150 mixer inlet and set it aside Tip You can use the tubing as the inlet tubing to the large volume gradient mixer However the existing tubing that connects the GM150 mixer outlet to the reference valve is too short to be used with the large volume mixer b Remove the tubing...

Page 64: ...le phases for operating principles see page 1 4 The degasser s vacuum system provides vacuum at a preset level to the connected vacuum chambers The vacuum system consists of the vacuum pump vacuum sensor and vacuum chambers The control system consists of the control board used to monitor and control the vacuum pump and the two color front panel LED see page 4 6 The degasser removes dissolved gasse...

Page 65: ...zel startup kit or 1 16 inch OD stainless steel Four ferrules and compression screws startup kit for each channel Tubing 0 149 inch OD PTFE 1 startup kit Connecting tubing to the degasser inlet and outlet The connectors on the front panel of the degasser are 1 4 28 flat bottom fittings The startup kit contains the compression screws and ferrules needed to connect tubing to these fittings 1 Polytet...

Page 66: ...nlet and outlet fittings to prevent leaks Tip To minimize degasser equilibration time when using only one pump fill the degas tubing in the unused vacuum chamber with a fluid such as water or eluent Install caps on the chamber s inlet and outlet fittings to prevent leaks Installing the degasser vent line Besides removing dissolved gases from the eluents the degasser can vaporize some eluent compon...

Page 67: ...ally while you perform HPLC runs There are no controls to adjust as it removes gases from the eluents For additional information see page 4 2 Installing the plunger seal wash system The plunger seal wash system is an optional accessory for all of the 1500 series pumps The seal wash solvent lubricates the plunger and flushes away any solvent or dried salts forced past the plunger seal from the high...

Page 68: ... inch TORX wrench T15 HPLC grade methanol Tweezers or tape Razor knife or tubing cutter Preparing the instrument To prepare the pump 1 Purge the pump with methanol see Chapter 4 If methanol is not miscible with your mobile phase use an intermediate mobile phase 2 Insert the priming syringe into the Luer fitting at the center of the draw off valve handle and then rotate the handle counterclockwise ...

Page 69: ...t rest on the plunger as you remove the head 3 Loosen and remove the inlet and outlet tubing assemblies from the pump head For 1515 1525 pump use a 5 16 inch open end wrench to hold the knurled portion of the check valve housings in place while loosening the inlet and outlet tubing assemblies with another 5 16 inch open end wrench see the figure below For the 1525EF and 1525µ pump use the 1 2 inch...

Page 70: ...n page 3 27 If you are installing the seal wash kit in a 1515 1525 or 1525EF model continue with step 7 below 7 While holding the head support in place use the 9 64 inch T handle hex wrench to remove the four head support mounting screws 8 Carefully slide the pump head support straight off 9 Complete the procedure below Installing head support components 1515 1525 pump only or page 3 25 as appropr...

Page 71: ... back side of the pump head support by its larger wash seal cavity The following figure shows the back side of the pump head support Pump head support components 1515 1525 pump 2 Insert the wash seal spacer into the wash seal cavity in the center of the head support Tip Because the spacer fits tightly start its edge into the cavity at a slight angle and then rotate it into the cavity while pressin...

Page 72: ...ght source and looking through the holes in the edge of the head support The fluid path must be unobstructed If it is not blow through the hole or use a thin wire to clear any obstructions 6 Lay the head support on a hard surface back side up 7 Insert the plunger wash seal spring side down into the wash seal cavity in the center of the head support 8 Use the wash seal installation tool to bottom t...

Page 73: ...wash fitting installation tool to start the retaining ring into the wash seal cavity 13 Carefully remove the wash fitting installation tool and then use the wash seal installation tool to bottom the retaining ring against the wash seal 14 Insert a tube fitting barbed end out into each hole on the edge of the head support 15 Slide the tubular end of the wash fitting installation tool over the fitti...

Page 74: ... and then slide the head onto the head support 23 Insert the two 5 32 inch hex screws and alternately tighten them with a T handle wrench 24 Ensure the gap between the pump head and the head support is uniformly spaced all around 25 Pull out and release the indicator rod If the rod does not snap back easily the head is misaligned Loosen the pump head and then repeat step 21 through step 23 above I...

Page 75: ...The following figure shows the back side of the pump head support 1525EF pump head support components 2 Insert the wash seal spacer into the wash seal cavity at the center of the head support 3 Using the wash seal installation tool press the wash seal spacer until it bottoms in the wash seal cavity 4 Rotate the wash seal spacer with a screwdriver until its slots line up with the holes in the edge ...

Page 76: ...m the plunger wash seal against the wash seal spacer 9 Insert a tube fitting barbed end out into each hole on the edge of the head support 10 Slide the tubular end of the wash fitting installation tool over the fitting and press down until the fitting flange is firmly seated against the flat area on the head support edge 11 Slide the stainless steel washer spring and plastic washer included in the...

Page 77: ...ease the indicator rod If the rod does not snap back easily the head is misaligned Loosen the pump head and then repeat step 17 through step 19 If the rod snaps back easily continue with step 21 22 Reconnect the inlet and outlet tubing assemblies to the pump head 23 Use the 1 2 inch open end wrench to hold the check valve housings in place while tightening the tubings with the 5 16 inch open end w...

Page 78: ...es the seal retainer see the figure above 4 Carefully slide the retainer assembly over the plunger and into the head support 5 Ensure the indicator rod slides freely 6 If the pump head has a plastic cap behind the top check valve housing use the 5 16 inch open end wrench to remove the cap 7 Carefully start the barbed fittings into the threaded holes at the top and bottom of the pump head Tighten t...

Page 79: ... in place while tightening the tubings with the 5 16 inch open end wrench 13 Repeat the process for the remaining pump heads and then complete the steps in Installing the solenoid below Installing the solenoid Perform these steps for all 1500 series pumps 1 Install the solenoid bracket on the pump chassis with the two TORX head screws provided with the kit Tip The solenoid bracket mounting holes a...

Page 80: ...de of the solenoid base and tighten the fitting finger tight 6 Plug the solenoid cable into the solenoid cable connector The solenoid cable must be plugged in at all times to ensure that the seal wash system is active whenever the instrument is operating 7 Affix the warning sticker to the pulse dampener shroud 8 For all models complete Installing seal wash tubing on page 3 31 Positioning arrow Sol...

Page 81: ...ing on the upper right pump head 4 Connect one end of the 41 0 cm piece of tubing to the top fitting on the upper right pump head Connect the other end of the tube to the bottom fitting on the lower right pump head 5 Connect one end of the remaining 20 0 cm piece of tubing to the top fitting on the lower right pump head Connect the other end of the tube to the bottom fitting on the lower left pump...

Page 82: ...a compression screw over the end of the tubing followed by a ferrule with its tapered end facing away from the tubing end and its wide end flush with the tubing end as shown in the figure below Reverse ferrule and compression screw assembly IN OUT 41 0 cm 8 0 cm 20 0 cm 20 0 cm Upper left pump head Lower left pump head Upper right pump head Lower right pump head Seal wash waste tubing Seal wash in...

Page 83: ...bing into the seal wash solvent reservoir 12 Prime the pump and check for leaks Using the seal wash system The seal wash solvent lubricates the plunger It also flushes away solvent or precipitated salts forced past the plunger seal from the high pressure side of the solvent piston chamber Caution To avoid damaging the ferrule do not overtighten the compression screw Caution Once you install the se...

Page 84: ...3 34 Installing Options and Accessories ...

Page 85: ...0 series pump for making chromatographic runs via Waters data control software Contents Topic Page Startup and initial preparation 4 2 Preparing for Breeze 2 operation 4 10 Preparing for Empower 2 operation 4 15 Preparing for MassLynx operation 4 19 Powering off the pump 4 23 ...

Page 86: ...tem devices before you power on the system workstation and startup the data control application Ensure the pump is installed according to the instructions in Chapter 2 Ensure the options and accessories included with your pump are installed according to the instructions in Chapter 3 To power on the pump locate the power switch on the left hand side panel and set it to the 1 ON position Pump prepar...

Page 87: ...to obtain accurate reproducible chromatography To prevent air from being drawn into the pump keep the eluent level in the reservoir at least four inches 10 cm higher than the inlet manifold Do not place eluent reservoirs on top of a 1500 series pump Use only filtered degassed eluents After use with a buffer purge the pump with HPLC grade water When possible use eluents of similar viscosity when yo...

Page 88: ...s necessary during initial pump startup or when the entire eluent line is dry Tip For binary pumps you can possibly need to dry prime one or both pump assemblies If your pump configuration includes a plunger seal wash system you can possibly need to prime the seal wash pump as well see page 4 9 Required material Priming syringe P N WAT010337 in startup kit To dry prime the pump 1 Fully depress the...

Page 89: ...inary pumps repeat step 1 through step 5 to dry prime the second pump assembly if necessary Operating with the integral vacuum degasser If your HPLC pump includes the integral vacuum degasser electrical power is supplied to the degasser when you power on the pump With the pump s switch set to On the degasser operates automatically while you perform HPLC runs There are no controls to adjust as it r...

Page 90: ... The LED on the 1500 series pump front panel illuminates The vacuum pump begins its evacuation Normal operation begins when the vacuum pump reaches its vacuum target and changes from high to low RPM If the degasser fails you can continue pump operation without the degasser by setting the degasser toggle switch located at the top of the degasser unit to Off Tip The degasser is intended for use at a...

Page 91: ...g initial evacuation Steady green Degasser is operating with pump at low RPM vacuum level below 60 mmHgA 1 16 psiA normal operation Green flash 0 5 seconds on and 0 5 seconds off Degasser is operating but vacuum level is unstable This condition can also indicate a sudden change in degasser work load Yellow flash 0 5 seconds on and 2 seconds off Degasser is not operating Degasser toggle switch is s...

Page 92: ...seal wash pump then shifts to its normal operating mode intermittently pumping seal wash solvent according to the duty cycles in the following table Once you stop using the instrument seal washing continues for 30 minutes after the mobile phase stops flowing Tip The seal wash pump returns to the priming sequence each time the instrument is powered on However the seal wash pump requires priming onl...

Page 93: ...precipitation problems when you switch between GPC and reversed phase analytical solvents Priming the plunger seal wash pump You must prime the plunger seal wash pump for first time use or when the entire line is dry Ensure the plunger seal wash supply line labeled Seal Wash In is in the plunger seal wash solvent bottle and that the plunger seal wash waste line clear is in an appropriate waste con...

Page 94: ...ntrol software do not exceed the maximum flow rate that is specified for the pump For the 1515 or 1525 pump combined flow from Pumps A and B do not exceed 10 00 mL minute For the 1525EF combined flow from Pumps A and B do not exceed 22 5 mL minute For the 1525µ pump combined flow from Pumps A and B do not exceed 5 00 mL minute Tip The data control software can possibly allow you to enter values th...

Page 95: ...ted to waste bypassing any connected components downstream of the pump by turning the pump s reference valve knob 1 2 turn to the right for the 1525µ pump position the vent valve knob to Vent To prime and purge the pump 1 At the Breeze 2 workstation log in to Breeze 2 2 In the Select Project and System dialog box verify that your chromatographic system with a 1500 series pump is configured and onl...

Page 96: ...llow the pump to run for approximately one minute and then verify that the eluent flow exiting to waste is normal and consistent for the specified flow rate 6 Click the Stop Flow icon in the Run Samples window to stop the flow Purging the flow path via Breeze 2 control Purging ensures that all eluent in the flow path is replaced with new eluent before you equilibrate the system and run samples see...

Page 97: ...your column and application and then follow step 1 through step 4 on page 4 11 2 Allow the pump to run for a few minutes to complete the purge Tip The amount of running time required to purge the flow path depends on total system volume eluent miscibility flow rate and so on 3 Verify that the flow exiting to waste is normal and consistent for the specified flow rate and then click the Stop Flow ic...

Page 98: ... 4 11 2 Allow the system to equilibrate for several minutes Tip The amount of running time required for equilibration depends on total system volume eluent miscibility flow rate and so on 3 Monitor the system pressure to verify it is normal and stable 4 Inspect the eluent flow exiting the system to verify it is normal and consistent for the specified flow rate Tip To monitor the detector baseline ...

Page 99: ...you power on the Empower 2 system workstation and start the Empower 2 application see page 4 2 Priming and purging the pump via Empower 2 control You must prime and purge the pump to ensure its proper operation see page 4 2 Before priming and purging the pump If the pump is dry dry prime it before proceeding see page 4 4 Ensure the eluent flow is directed to waste bypassing any connected component...

Page 100: ...low rate and gradient settings Tip To quickly prime and purge just the pump for the 1515 pump set the rate to 2 0 mL min for the 1525 pump set the rate to 4 0 mL min and gradient to 50 A 50 B for the 1525µ pump set the rate to 2 0 mL min and gradient to 50 A 50 B for the 1525EF pump set the rate to 6 0 mL min and gradient to 50 A 50 B 7 Select Immediate Flow Change and click OK 8 Allow the pump to...

Page 101: ...miscible perform the steps in this procedure with an intermediate eluent before purging with the final eluent See Appendix C for more information Purging To purge the flow path 1 Determine flow rate and gradient settings appropriate for your column and application and then follow step 1 through step 7 on page 4 15 2 Allow the pump to run for a few minutes to complete the purge Tip The amount of ru...

Page 102: ...mine flow rate and gradient settings appropriate for your column and application and then follow step 1 through step 7 on page 4 15 2 Allow the system to equilibrate for several minutes Tip The amount of running time required for equilibration depends on total system volume eluent miscibility flow rate and so on 3 Monitor the system pressure to verify it is normal and stable Inspect the eluent flo...

Page 103: ...hic system devices before you power on the MassLynx system workstation and start the MassLynx application see page 4 2 Priming and purging the pump via MassLynx control You must prime and purge the pump to ensure its proper operation see page 4 2 Before priming and purging the pump If the pump is dry dry prime it before proceeding see page 4 4 Ensure the eluent flow is directed to waste bypassing ...

Page 104: ...50 B for the 1525EF pump set the rate to 6 0 mL min and gradient to 50 A 50 B 6 Click the Load Method icon to download the method to the system Doing so initiates the pump and starts flow 7 Allow the pump to run for approximately one minute and then verify that the eluent flow exiting to waste is normal and consistent for the selected flow rate 8 Click the Start Stop Pumping icon in the Inlet Meth...

Page 105: ... purge the flow path 1 Determine flow rate and gradient settings appropriate for your column and application and then follow step 1 through step 6 on page 4 19 2 Allow the pump to run for a few minutes to complete the purge Tip The amount of running time required to purge the flow path depends on total system volume eluent miscibility flow rate and so on 3 Verify that the flow exiting to waste is ...

Page 106: ...Vent To equilibrate the system 1 Determine flow rate and gradient settings appropriate for your column and application and then follow step 1 through step 6 on page 4 19 2 Allow the system to equilibrate for several minutes Tip The amount of running time required to equilibrate the system depends on total system volume eluent miscibility flow rate and so on 3 Monitor the system pressure and detect...

Page 107: ...idle for more than one day purge with a methanol water solution to prevent the growth of microorganisms 3 If the pump is running use a shutdown method to ramp the flow to zero 4 Turn the power switch to the O OFF position Caution Since abrupt flow changes can damage columns it is advisable to ramp flow to 0 using an appropriate shutdown method Refer to your data control software s online Help for ...

Page 108: ...4 24 Preparing for Operation ...

Page 109: ...ribes how to run diagnostic tests and set pump calibration parameters and explains how to replace pump components Contents Topic Page Maintenance considerations 5 2 Performing pump diagnostic tests 5 3 Replacing and cleaning plunger seals and plungers 5 6 Replacing check valves 5 14 Replacing a draw off valve 5 22 Replacing fuses 5 25 ...

Page 110: ...nter any problems replacing parts in the 1500 series pump contact Waters Technical Service Warning To prevent injury always observe Good Laboratory Practices when you handle eluents change tubing or operate the 1500 series HPLC pump Know the physical and chemical properties of the eluents Refer to the Material Safety Data Sheets for the eluents in use To avoid the possible eye injury or cuts handl...

Page 111: ...er seals air bubbles in the lines incorrectly set flow rate leaks or other pump related problems Be aware that other factors such as system and column equilibration column age operating temperature and so on can also affect retention time stability See your system suitability documentation for more information Ramp and decay test Use the ramp and decay test to monitor check valve performance Requi...

Page 112: ...losed and the eluent directed to waste by turning the reference valve 1 2 turn to the right for 1525μ position the vent valve to vent 7 Use the data control software to set the following pump parameter values Flow rate For 1515 isocratic and 1525 binary pump 0 3 mL min For 1525EF pump 0 5 mL min For 1525µ pump 0 2 mL min High pressure limit For 1515 isocratic 1525 binary pump and 1525µ pump 41 370...

Page 113: ...ng the reference valve knob 1 2 turn to the left for the 1525µ position the vent valve to the system 12 Disconnect the right hand pump head outlet tubing from the draw off valve 13 Transfer the compression plug from the left hand port of the draw off valve to the right hand one 14 Reconnect the left hand pump head outlet tubing to the left hand port of the draw off valve 15 Repeat step 5 through s...

Page 114: ...ance operation of the pump perform the following maintenance tasks Replace the plunger seals and bearings twice yearly or as needed Replace the plunger twice yearly or as needed Clean and inspect the plunger every six months or sooner if using abrasive eluents Preparing for plunger seal replacement Required materials Open end wrench 5 16 inch startup kit Allen wrench 5 32 inch startup kit Adjustab...

Page 115: ...methanol 4 Use the data control software to set the flow rate to 0 3 mL min and to initiate pump flow 5 When the indicator rod fully retracts into the pump head use the software to stop the flow turning off the pump Tip This action ensures that the weight of the pump head does not rest on the plunger while you remove the head 6 While using the adjustable wrench to hold the check valve housings in ...

Page 116: ...nd of the tool clockwise into the seal and then pull the seal to remove it from the pump head 3 Place the new plunger seal onto the tip of the seal insertion tool Tip Make sure the spring in the seal is facing away from the end of the tool you must be able to see the seal spring at the tip of the tool 4 Moisten the seal with methanol 5 Use the seal insertion tool to firmly seat the seal in the pum...

Page 117: ...nd then use the software to stop the flow turning off the pump 15 Use the adjustable wrench to hold the check valve housings in place while reconnecting the inlet and outlet tubing to the pump head 16 Tighten the tubing using the 5 16 inch open end wrench 17 Prime the pump as described in Chapter 4 If you notice leaks verify pump head and plunger seal installation Cleaning and replacing the plunge...

Page 118: ...ment plunger Replacement plunger seals Removing the plunger To remove the plunger from the pump 1 Remove the pump head as described on page 5 6 2 Remove the four head support screws with the 9 64 inch Allen wrench and then carefully slide the head support assembly and the indicator rod off the pump Exposed head support assembly 3 Set the head support assembly on the benchtop 4 Use the data control...

Page 119: ... Carefully remove the plunger For a 1525µ pump slide the plunger assembly out of the back of the support assembly Set it aside for cleaning For 1515 1525 pumps use the snap ring pliers to remove the snap ring that holds the plunger in place see the figure below Carefully remove the plunger assembly and set it aside for cleaning Removing the plunger assembly 1515 1525 pumps ...

Page 120: ...hem with your fingers If the plunger is not scratched or otherwise damaged reassemble it with new seals Continue with Replacing the Plunger on page 5 12 If the plunger is damaged replace both the plunger and the seals Continue with Replacing the Plunger on page 5 12 Replacing the Plunger To replace the plunger 1 Reassemble the components of the plunger assembly as shown in the figure above 2 Lubri...

Page 121: ... seated in the groove before you release the pliers Inserting the plunger 1515 1525 pumps 4 Use the data control software to set the flow rate to 1 0 mL min and initiate pump flow 5 When the indicator rod is fully retracted into the pump head use the software to stop the flow turning off the pump 6 With the indicator rod hole oriented to the upper right to the upper left for 1525µ reinstall the he...

Page 122: ...en repeat step 8 through step 11 If the rod moves smoothly snaps back easily use the software to stop the flow turning off the pump Continue with step 13 13 Using the adjustable wrench to hold the check valve housings in place reconnect the inlet and outlet tubing assemblies to the pump head Tighten the tubings using the 5 16 inch open end wrench 14 Prime the pump as described in Chapter 4 If you ...

Page 123: ...stable wrench 1 2 inch startup kit Priming syringe startup kit Replacement check valves assembled with housings HPLC grade methanol Forceps Replacing 1515 1525 check valves The 1515 isocratic and 1525 binary pumps employ a check valve cartridge for both the inlet and outlet check valves To replace a check valve 1 Purge the pump with methanol see Chapter 4 If methanol is not miscible with your curr...

Page 124: ...The arrow printed on the check valve cartridge indicates the direction that liquid can flow For both inlet and outlet check valve assemblies the arrow must point upward when installed in the pump head To help keep the cartridge seated within the housing moisten it with methanol during installation Outlet check valve assembly 1515 1525 pumps Check valve cartridge Check valve housing ...

Page 125: ...e with your current eluent use an intermediate eluent 2 While holding the inlet check valve housing with the adjustable wrench use the 5 16 inch open end wrench to loosen the compression screw and remove the tubing assembly from the check valve housing see the figure below 3 Use the adjustable wrench to remove the entire check valve assembly including the housing from the pump head 4 Replace the e...

Page 126: ... rest on the plunger when you remove the pump head 3 Use the adjustable wrench to hold the housings in place Use the 5 16 inch open end wrench to loosen the compression screws and remove the inlet and outlet tubing assemblies from the check valve housings Disconnecting the outlet tubing 1525EF pump 4 Use the adjustable wrench to loosen the outlet check valve housing 5 While holding the pump head i...

Page 127: ... 12 Reinstall the outlet tubing and inlet tubing assemblies Tip Use the 5 16 inch open end wrench to tighten the compression screw while holding the housing with the adjustable wrench 13 Prime the pump as described in Chapter 4 Check for leaks Replacing 1525µ check valves This section describes how to replace inlet outlet and restrictor tee check valves for the 1525µ pump These check valves employ...

Page 128: ...roper operation the arrow must point upward when installed on the pump Tip To help keep the cartridge seated within the housing moisten it with methanol during installation 5 With the housing held upright to prevent the cartridge from falling out finger tighten the housing into the pump head Tighten the check valve another 1 4 turn with the adjustable wrench 6 Reinstall the inlet tubing assembly T...

Page 129: ...tubing assemblies from the check valve housings 4 Use the adjustable wrench to loosen the outlet check valve housing 5 While holding the pump head securely against the pump use the 5 32 inch Allen wrench to remove the two pump head assembly mounting screws Loosen the screws 1 2 turn at a time for the first two turns 6 Carefully slide the pump head assembly off the pump see the instructions on page...

Page 130: ...sion screw while holding the housing with the adjustable wrench 13 Reinstall the inlet tubing assembly Tip Use the 5 16 inch open end wrench to tighten the compression screw while holding the housing with the adjustable wrench 14 Prime the pump as described in Chapter 4 Check for leaks Replacing the 1525µ restrictor tee check valves To replace the restrictor tee check valves follow the procedure o...

Page 131: ...t gravity flow of eluent Disconnect the eluent supply line from the inlet manifold on the pump 2 Insert the priming syringe into the Luer fitting at the center of the draw off valve handle and then rotate the handle counterclockwise about 1 2 turn to open the valve 3 Use the syringe to withdraw all the methanol 4 Use the 5 16 inch open end wrench to disconnect the three stainless steel compression...

Page 132: ... install the draw off valve 1 Attach the new draw off valve to the bracket using the two Phillips screws Tip Ensure the new valve is oriented on the bracket as shown in the figure above 2 Reposition the draw off valve inlet manifold assembly onto the pump being careful not to pinch the stainless tubing behind the bracket When correctly positioned the stainless tubing extends through the cutouts on...

Page 133: ...nel fuse holder Identifying faulty fuses Suspect a faulty fuse if either of these conditions occur The pump fails to power on The fan does not operate Warning To avoid possible electrical shock power off and unplug the instrument before checking the fuses For continued protection against fire hazard replace fuses only with those of the same type and rating Fuse holder ...

Page 134: ...ulty fuse Power off the pump and then remove the power cord from its connector Use the small flat blade screwdriver to remove the fuse holder located just above the power cord connector 1 Remove and discard the old fuses 2 Insert the new fuses into the fuse holder 3 Insert the fuse holder into the receptacle and gently push until it locks into position ...

Page 135: ...identify and correct noises and identify pump related chromatographic problems Warning To prevent injury always observe Good Laboratory Practices when you troubleshoot the 1500 series pump Contents Topic Page Troubleshooting pump problems 6 2 Identifying and correcting noises 6 8 Identifying chromatographic problems 6 9 ...

Page 136: ...imit to the correct value Blocked column Clean the column according to the column care and use manual Blocked mixer Backflush or disassemble the mixer Viscous eluents in use Verify eluent viscosity the observed pressure may be normal for the column eluent blend If necessary change to a less viscous eluent Defective pressure transducer Contact Waters Technical Service Blocked outlet tubing or fluid...

Page 137: ...perly connected to power source and pump No power at outlet Confirm the outlet is serviceable by connecting to another electrical unit known to be working If that unit does not work relocate the pump to a functioning electrical outlet Blown fuse Replace the fuses Pump not delivering eluent Draw off valve open or leaking Close the draw off valve If eluent still leaks replace the valve see page 5 22...

Page 138: ...ening and then retightening the check valve to dislodge any stuck particles Often a sticking check valve is unstuck by letting the pump run for a few minutes at pressure If there is no flow from one head replace the head s inlet check valve see page 5 14 Damaged plunger seal indicated by eluent leaking from behind the pump head or by salt crystal build up around the back of the pump head Confirm t...

Page 139: ... head Tubing ID too small Perform the following corrective action Raise eluent reservoirs above the pump Inspect tubing Tighten straighten or replace tubing Verify degasser operation Replace filter Prime pump see Chapter 4 Use correct tubing Ruptured high pressure noise filter Contact Waters Technical Service Defective pump motor Contact Waters Technical Service Defective circuit board Contact Wat...

Page 140: ... draw off valve Draw off valve open or broken Close the draw off valve If the leak continues replace the valve see page 5 22 Erratic flow rate pump pulsations Pump not primed Prime the pump If you are using a volatile eluent such as hexane prime the pump with a miscible less volatile eluent such as THF or methanol Ensure that the column is disconnected to avoid disrupting equilibrium Reservoir low...

Page 141: ...for blockages Replace the inlet filter frit Pump plunger seal leaking under pump head Replace pump plunger seal see page 5 6 Worn pump plunger Replace the plunger see page 5 6 Immiscible eluents in pump head Refer to the Corrective Action under the Pump not delivering eluent symptom listed above Pump cavitation Refer to the Corrective Action s under the Pump not delivering eluent symptom listed ab...

Page 142: ...unger seal If click does not stop and cannot be isolated to one pump head replace the plunger seals one at a time see page 5 6 Worn indicator rod spring Replace the spring Squeak Plunger seals dry Wet plunger with appropriate eluent through pump head access holes Binding plunger seal Replace plunger seal assembly see page 5 6 Improper plunger seal Install correct pump plunger seal see page 5 6 Bin...

Page 143: ... valves see page 5 14 Leaking pump seals Replace pump seals see page 5 6 Leaks around fittings Tighten fittings Separation chemistry Check eluent and column Clogged eluent filters Replace filters Increased retention times Incorrect flow rate Change flow rate Incorrect eluent composition Change eluent composition Column heater not turned on Turn column heater on Incorrect eluent Use correct eluent ...

Page 144: ...sed Degas eluent Incorrect chemistry integration Verify chemistry integration Baseline drift rapid Column not equilibrated Equilibrate column Eluent contaminated Use fresh eluent Eluent not properly degassed rapid or slow drift Degas eluent Flow fluctuations rapid or slow drift Fix pump problems replace pump seals and check valves Baseline drift slow Eluent contaminated Use fresh eluent Ambient te...

Page 145: ...erm cycling Disconnect other instruments on the power line try a different wall outlet have line voltage checked use power conditioner Radio frequency noise short or long term cycling Eliminate interference Baseline noise cycling long term for 1 hour Ambient temperature fluctuations Stabilize ambient temperature Integrator or recorder faulty Inspect integrator or recorder for excessive baseline no...

Page 146: ...onditioner Recorder voltage incorrect Set recorder to correct voltage Radio frequency noise Eliminate interference Straight baseline no peaks No pump flow Set pump flow rate Leak in eluent path Repair the leak Sensitivity loss Degraded contaminated or improperly prepared sample Use fresh sample Column contaminated Clean or replace column Loss of column efficiency Clean or replace column Change in ...

Page 147: ...ides also include the hazard symbols with accompanying text statements describing the hazards and telling you how to avoid them This appendix presents all the safety symbols and statements that apply to the entire line of Waters products Contents Topic Page Warning symbols A 2 Caution symbol A 5 Warnings that apply to all Waters instruments A 6 Electrical and handling symbols A 12 ...

Page 148: ... electric shocks ultraviolet radiation exposures and others When the following symbols appear in a manual s narratives or procedures their accompanying text identifies the specific risk and explains how to avoid it Warning General risk of danger When this symbol appears on an instrument consult the instrument s user documentation for important safety related information before you use the instrume...

Page 149: ...ning Risk of explosion Warning Risk of eye injury Warning Pressurized nonmetallic or polymer tubing can burst Observe these precautions when working around such tubing Wear eye protection Extinguish all nearby flames Do not use tubing that is or has been stressed or kinked Do not expose nonmetallic tubing to incompatible compounds like tetrahydrofuran THF and nitric or sulfuric acids Be aware that...

Page 150: ... Ensure that the nitrogen supply pressure never falls below 690 kPa 6 9 bar 100 psi during an analysis in which flammable solvents are used Also ensure a gas fail connection is connected to the LC system so that the LC solvent flow stops if the nitrogen supply fails Warning To avoid electric shock do not remove the mass spectrometer s protective panels The components they cover are not user servic...

Page 151: ...anisms and other biological materials To avoid infection with these agents assume that all biological fluids are infectious observe Good Laboratory Practices and consult your organization s biohazard safety representative regarding their proper use and handling Specific precautions appear in the latest edition of the US National Institutes of Health NIH publication Biosafety in Microbiological and...

Page 152: ...e ausdrückliche Genehmigung der für die ordnungsgemäße Funktionstüchtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems führen Avvertenza qualsiasi modifica o alterazione apportata a questa unità e non espressamente autorizzata dai responsabili per la conformità fa decadere il diritto all utilizzo dell apparecchiatura da parte dell utente Atencion cualquier cambio ...

Page 153: ...e trouvant à proximité de l instrument Evitez d utiliser des tubes sévèrement déformés ou endommagés Evitez d utiliser des tubes non métalliques avec du tétrahydrofurane THF ou de l acide sulfurique ou nitrique concentré Sachez que le chlorure de méthylène et le diméthylesulfoxyde entraînent le gonflement des tuyaux non métalliques ce qui réduit considérablement leur pression de rupture Vorsicht B...

Page 154: ...ottura dei tubi stessi Advertencia se recomienda precaución cuando se trabaje con tubos de polímero sometidos a presión El usuario deberá protegerse siempre los ojos cuando trabaje cerca de tubos de polímero sometidos a presión Si hubiera alguna llama las proximidades No se debe trabajar con tubos que se hayan doblado o sometido a altas presiones Es necesario utilizar tubos de metal cuando se trab...

Page 155: ...의 화기를 모두 끄십시오 심하게 변형되거나 꼬인 튜브는 사용하지 마십시오 비금속 Nonmetallic 튜브를 테트라히드로푸란 Tetrahydrofuran THF 또는 농축 질산 또는 황산과 함께 사용하지 마십시오 염화 메틸렌 Methylene chloride 및 디메틸술폭시드 Dimethyl sulfoxide 는 비금속 튜브를 부풀려 튜브의 파열 압력을 크게 감소시킬 수 있으므로 유의하십시오 警告 圧力のかかったポリマーチューブを扱うときは 注意してください 加圧されたポリマーチューブの付近では 必ず保護メガネを着用してください 近くにある火を消してください 著しく変形した または折れ曲がったチューブは使用しないでください 非金属チューブには テトラヒドロフラン THF や高濃度の硝酸または硫酸などを流 さないでください 塩化メチレンやジメチル...

Page 156: ...rheitseinrichtungen unter Umständen nicht ordnungsgemäß funktionieren Attenzione si rende noto all utente che l eventuale utilizzo dell apparecchiatura secondo modalità non previste dal produttore può compromettere la protezione offerta dall apparecchiatura Advertencia el usuario deberá saber que si el equipo se utiliza de forma distinta a la especificada por el fabricante las medidas de protecció...

Page 157: ...nwert auf den Tafeln neben den Sicherungsabdeckungen des Geräts gedruckt sind Attenzione per garantire protezione contro gli incendi sostituire i fusibili con altri dello stesso tipo aventi le caratteristiche indicate sui pannelli adiacenti alla copertura fusibili dello strumento Advertencia Para evitar incendios sustituir los fusibles por aquellos del tipo y características impresos en los panele...

Page 158: ...se can appear in instrument user manuals and on the instrument s front or rear panels Electrical power on Electrical power off Standby Direct current Alternating current Protective conductor terminal Frame or chassis terminal Fuse Recycle symbol Do not dispose in municipal waste ...

Page 159: ...ing symbols A 13 Handling symbols These handling symbols and their associated text can appear on labels affixed to the outer packaging of Waters instrument and component shipments Keep upright Keep dry Fragile Use no hooks ...

Page 160: ...A 14 Safety Advisories ...

Page 161: ...egasser Instrument control and communication Physical specifications for 1500 series pump Item Specification Height Depth Width Weight 1525 Weight 1515 43 cm 61 0 cm 30 5 cm without bottle holder 27 2 kg 20 4 kg Wetted Surface Material 316 stainless steel sapphire reinforced fluorocarbon polymer seals 1525µ pump UHMWPE seals 1525EF pump carbon reinforced Tefzel ...

Page 162: ...Audible Noise 70 dBA at operator position Solvent Compatibility Solvents consistent with materials of construction Salts and buffers can reduce seal life especially at pressures in excess of 20 684 3 kPa 206 8 bar 3000 psi Electrical specifications Item Specification Protection classa Class I Overvoltage categoryb Category II Pollution degree c Degree 2 Moisture protection d Normal IPXO Power requ...

Page 163: ...porary conductivity caused by condensation must be expected d Moisture Protection Normal IPXO IPXO means that there is no Ingress Protection against any type of dripping or sprayed water The X is a placeholder to identify protection against dust if applicable Performance specifications for the 1515 and 1525 pumps Item Specification Programmable Flow Rate Range 100 µL heads 0 00 to 10 00 mL min in ...

Page 164: ...ith 50 µL heads Pressure ripple one pump 2 0 at 0 5 mL min degassed methanol at 6894 75 kPa 68 9 bar 1000 psi to 13 789 51 kPa 137 9 bar 2000 psi backpressure Flow precision 2 seconds SD six replicates based on retention time or volumetric measures at 0 2 mL min 6894 75 kPa 68 9 bar 1000 psi to 13 789 51 kPa 137 9 bar 2000 psi backpressure PQ test each pump individually Flow accuracy 2 of setting ...

Page 165: ...ressure ripple one pump 3 0 at 2 25 mL min degassed methanol at 6894 75 kPa 68 9 bar 1000 psi backpressure Flow precision 0 1 RSD or 2 seconds SD six replicates based on retention time or volumetric measures at 1 mL min 6894 75 kPa 68 9 bar 1000 psi to 13 789 51 kPa 137 9 bar 2000 psi backpressure PQ test each pump individually Flow accuracy 1 0 of setting at 1 mL min or 30 µL min whichever is lar...

Page 166: ...al injector and no column heater Integral vacuum degasser operational specifications Item Specification Chemical resistance Unaffected by full range of organic solvents and aqueous solutions of acids bases salts and surfactants pH range 0 to 14 Gas removal efficiency Varies with flow rate see the table above Typical operating flow range 200 µL min to 10 mL min 1525 model 200 µL min to 5 mL min 152...

Page 167: ... inside diameter ID 1 8 meter tubing Vacuum source Built in 2 head diaphragm pump solvent resistant Vacuum sensor Detects vacuum from 0 to 15 6 psiA Maximum pressure on inlet and outlets 70 kPa 0 7 bar 10 psi a High density polyethylene b polytetrafluoroethylene c ethylenetetrafluoroethylene Teflon AF chemical compatibility test results Compatiblea Incompatibleb Acetonitrile Fluorinert Chloroform ...

Page 168: ...ght of Teflon AF after prolonged exposure at 23 C b Incompatible solvents immediately and irreversibly damage Teflon AF Instrument control and communication specifications Item Specification Inject Voltage Range 0 to 30 V Current 200 mA Stop Flow Input 0 to 30 V Contact Closure Outputs Voltage Range 30 V Current 500 mA External Control Ethernet or IEEE 488 Teflon AF chemical compatibility test res...

Page 169: ...siderations Contents Topic Page Introduction C 2 Solvent compatibility C 3 Solvent miscibility C 6 Buffered solvents C 8 Head height C 9 Solvent viscosity C 9 Mobile phase solvent degassing C 9 Wavelength selection C 12 ...

Page 170: ...ents through 0 45 µm filters before use Solvents distilled in glass generally maintain their purity from lot to lot use them to ensure the best possible results Preparation checklist The following solvent preparation guidelines help to ensure stable baselines and good resolution Filter solvents with a 0 45 µm filter Degas the solvent Stir the solvent Keep in a place free from drafts and shock Wate...

Page 171: ...s HPLC pump However long term static exposure to halide salts for example fluoride bromide chloride and iodide cause pitting and corrosion of stainless steel parts When using these salts flush your system thoroughly with water if the pump will be idle for more than two days Solvents to use Materials of construction used in a 1500 series HPLC pump do not react with most acids bases salts and organi...

Page 172: ...m acetate Trilithium citrate H2O2 up to 10 KHCO3 NaH2BO3 Tris HIBA KMnO4 NaHCO3 4 2 pyridylazo resorcinol mono sodium salt monohydrate K2CO3 KNO3 NaHSO4 K2Cr2O3 LiCl04 NaNO3 K2S Na2B4O7 NaOCl Acids for use with the 1500 series HPLC pump Acids Acetic acid glacial Hydrochloric acid Perchloric acid Benzoic acid Lactic acid Phosphoric acid Chromic acid Methanesulphonic acid Pyridine 2 6 dicarboxylic a...

Page 173: ...phenol Chloroform Ethylene glycol Methylene chloride Acetone Cyclohexane Formaldehyde n Propanol Acetonitrile Cyclohexanone Heptane Phenol Amyl acetate Dibutyl phathalate Hexane Tetrahydrofuran THF Benzaldehyde Dimethyl formamide iso Octane Toluene Benzene Dimethyl sulfoxide iso Propanol Waters PIC Reagents Benzyl alcohol Ethanol Lysine hydrochloride Xylene Butanol Ethyl acetate Methanol Carbon te...

Page 174: ...re on solvent solubility Buffers dissolved in water may precipitate when mixed with organic solvents When you switch from a strong buffer to an organic solvent flush the buffer out of the system with distilled water before you add the organic solvent Solvent miscibility Polarity index Solvent Viscosity CP 20 C Boiling point C 1 atm Miscibility number M λ Cutoff nm 0 3 N decane 0 92 174 1 29 0 4 Is...

Page 175: ... 20 4 6 Benzonitrile 1 22 191 1 15 19 4 8 Dioxane 1 54 101 3 17 220 5 2 Ethanol 1 20 78 3 14 210 5 3 Pyridine 0 94 115 3 16 305 5 3 Nitroethane 0 68 114 0 5 4 Acetone 0 32 56 3 15 17 330 5 5 Benzyl alcohol 5 80 205 5 13 5 7 Methoxyethanol 1 72 124 6 13 6 2 Acetonitrile 0 37 81 6 11 17 210 6 2 Acetic acid 1 26 117 9 14 6 4 Dimethylfor mamide 0 90 153 0 12 6 5 Dimethylsulfoxide 2 24 189 0 9 6 6 Meth...

Page 176: ...ates the degree of miscibility with highly lipophilic solvents The second number applies to the opposite end of the scale A large difference between these two numbers indicates a limited range of miscibility For example some fluorocarbons are immiscible with all the standard solvents and have M numbers of 0 and 32 Two liquids with dual M numbers are usually miscible with each other A liquid is cla...

Page 177: ...ges affect the analysis is not known monitor the pressure during the run Mobile phase solvent degassing Mobile phase difficulties account for the majority of all liquid chromatographic problems Using degassed solvents is important especially at wavelengths below 220 nm Degassing provides Stable baselines and enhanced sensitivity Reproducible retention times for eluting peaks Reproducible injection...

Page 178: ... the solubility of He in benzene increases with an increase in temperature Effects of partial pressure The mass of gas dissolved in a given volume of solvent is proportional to the partial pressure of the gas in the vapor phase of the solvent If you decrease the partial pressure of the gas the amount of that gas in solution also decreases Eluent degassing methods You can degas eluents using any of...

Page 179: ...s the solvent passes through the vacuum chamber At higher flow rates lesser amounts of gas per unit volume of solvent are removed Surface area of the degassing membrane The length of the degassing membrane is fixed in each vacuum chamber To increase the length of membrane you can connect two or more vacuum chambers in series Heating Heating to remove dissolved gas is usually not effective unless y...

Page 180: ...avelength at which the absorbance of the solvent is equal to 1 AU for some common chromatographic solvents Operating at a wavelength near or below the cutoff increases baseline noise due to the absorbance of the solvent UV cutoff wavelengths for common chromatographic solvents Solvent UV cutoff nm Solvent UV cutoff nm 1 Nitropropane 380 Ethylene glycol 210 2 Butoxyethanol 220 Iso octane 215 Aceton...

Page 181: ...5 Dioxane 215 Nitromethane 380 Ethanol 210 Petroleum ether 210 Ethyl acetate 256 Pyridine 330 Ethyl ether 220 Tetrahydrofuran 230 Ethyl sulfide 290 Toluene 285 Ethylene dichloride 230 Xylene 290 Wavelength cutoffs for different mobile phases Mobile phase UV cutoff nm Mobile phase UV cutoff nm Acetic acid 1 230 Sodium chloride 1 M 207 Ammonium acetate 10 mM 205 Sodium citrate 10 mM 225 Ammonium bic...

Page 182: ...0 215 Waters PIC Reagent B 6 1 vial liter 225 Potassium phosphate monobasic 10 mM dibasic 10 mM 190 190 Waters PIC Reagent B 6 low UV 1 vial liter 190 Sodium acetate 10 mM 205 Waters PIC Reagent D 4 1 vial liter 190 Refractive indices for common chromatographic solvents Solvent RI Solvent RI Fluoroalkanes 1 25 Tetrahydrofuran THF 1 408 Hexafluoroisopropanol HFIP 1 2752 Amyl alcohol 1 410 Methanol ...

Page 183: ... Ethylene dichloride 1 445 Isopropanol 1 38 Carbon tetrachloride 1 466 n Propanol 1 38 Dimethyl sulfoxide DMSO 1 477 Methylethylketone 1 381 Toluene 1 496 Diethyl amine 1 387 Xylene 1 50 n Propyl chloride 1 389 Benzene 1 501 Methylisobutylketone 1 394 Pyridine 1 510 Nitromethane 1 394 Chlorobenzene 1 525 1 Nitropropane 1 400 o Chlorophenol 1 547 Isooctane 1 404 Aniline 1 586 Cyclopentane 1 406 Car...

Page 184: ...C 16 Solvent Considerations ...

Page 185: ... performance 5 3 replacing 5 14 chemical compatibility test results B 7 chemical hazard warning A 5 chromatographic problems 6 9 cleaning plungers 5 12 column heater 1 10 components electronic 1 12 fluid handling 1 7 connecting eluent reservoir 2 11 power supply 2 5 pump inlet 2 12 connections electrical 2 4 guidelines 2 7 contacting Waters Technical Service 5 2 contamination 3 4 contamination elu...

Page 186: ...id waste lines connecting 2 16 fluid handling components 1 8 fuse faulty 5 25 holder 5 25 replacing 5 25 G gas solubility C 9 gradient mixer 1 10 guidelines connection 2 7 eluents 4 3 H handling symbols A 13 head height 4 3 6 5 heating eluents C 11 high pressure noise filter 6 5 high pressure shutdown 5 5 I identifying faulty fuses 5 25 IEEE 488 guidelines 2 7 setting device addresses 2 9 IEEE 488...

Page 187: ...ng 5 6 squeaking 6 8 worn 6 6 plungers inspecting 5 12 replacing 5 12 worn 6 6 power entry module 1 13 power supply connecting 2 5 power switch 1 13 powering off the pump 4 23 pressure transducer 1 10 6 3 6 7 pressure relieving 5 5 priming the pump 6 4 6 6 pulsations in eluent delivery 6 6 6 7 pulse dampeners 1 10 pump head assembly 1 10 pump head assembly removing 5 7 pump head support assembly r...

Page 188: ...bols caution A 5 electrical A 12 handling A 13 warning A 2 system pressure relieving 5 5 T tee 1 11 Teflon AF chemical compatibility test results B 7 tests ramp and decay 5 3 retention time stability test 5 3 THF tetrahydrofuran 6 6 C 3 troubleshooting chromatographic problems 6 9 noises 6 8 tubing 2 11 attaching compression fittings 2 15 cutting to length 2 15 eluent supply 4 4 making connections...

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